›› 2010, Vol. 53 ›› Issue (6): 647-654.doi:

• 研究论文 • 上一篇    下一篇

水稻褐飞虱内生共生细菌Arsenophonus的鉴定和系统分析

王渭霞,罗举,赖凤香,傅强   

  • 出版日期:2011-01-28 发布日期:2010-06-20
  • 通讯作者: 傅强

Identification and phylogenetic analysis of symbiotic bacteria Arsenophonus from the rice brown planthopper, Nilaparvata lugens (Stål) (Homoptera: Delphacidae)

WANG Wei-Xia, LUO Ju, LAI Feng-Xiang, FU Qiang   

  • Online:2011-01-28 Published:2010-06-20

摘要: 利用16S rDNA通用引物扩增了水稻褐飞虱Nilaparvata lugens(Stål)体内共生细菌的序列,经克隆、测序和NCBI数据库比对,发现褐飞虱体内存在杀雄菌属Arsenophonus类共生细菌,系统发育上与粉虱科和木虱科体内的Arsenophonus属亲源关系较近。在褐飞虱体内该共生细菌具有两种长度不同的16S rDNA序列,分别为1 504 bp和547 bp,其中后者为前者中间缺失了957 bp,其余序列相同。通过重新设计两对引物进行扩增,进一步确认不同褐飞虱地理种群及寄主种群均存在两种片段。Arsenophonus特异的 23S rDNA引物的扩增结果表明,Arsenophonus存在于所有检测的褐飞虱种群中,但不存在于水稻寄主中。荧光定量PCR检测发现3个褐飞虱室内寄主种群Arsenophonus属共生细菌含量不同,其中TN1种群明显高于Mudgo种群和ASD7种群。此为水稻褐飞虱体内存在Arsenophonus属共生细菌的首次报道。

关键词: 褐飞虱, 共生菌, 杀雄菌属, 16S rDNA, 系统发育

Abstract: PCR amplification of 16S rDNA with universal primers was conducted to detect the symbiotic bacteria of the rice brown planthopper, Nilaparvata lugens (Stål) . Analysis of the sequences showed that the brown planthoppers contain the symbiotic bacteria Arsenophonus,which has close relationship with Arsenophonus symbiont of Psyllidae and Aleyrodidae based on phylogenetic analyses. Two 16S rDNA sequences of Arsenophonus in N. lugens with different sizes were amplified, one is 1 504 bp and the other is 547 bp in length. The shorter one is the deleted product of the longer one by loss of its internal region of 957 bp. With two primers redesigned based on the above 16S rDNA sequences, the result of PCR amplification showed that the two different fragments existed in various host-associated populations and different geographic populations of N. lugens. Using 23S rDNA specific primers, the genus Arsenophonus was found in all the tested populations of N. lugens, but not in the host rice plant. The TN1 population carried significantly higher amount of Arsenophonus bacteria than the Mudgo population and ASD7 population assessed by quantitative real-time PCR. This is the first report of Arsenophonus bacteria from N. lugens.  

Key words: Nilaparvata lugens, symbiotic bacteria, Arsenophonus, 16S rDNA, phylogenetic relationship